Interactions between AR, ER, TR2, and TR4

Abstract

Disclosed are compositions and methods related to TR2, TR4, androgen receptor, and estrogen receptor and the interactions between these proteins.

What is claimed is:  
     
         1 . A composition comprising a fragment of TR2, wherein the composition interacts with ER, such that ER transcriptional activity is decreased relative to transcriptional activity in the absence of the composition.  
     
     
         2 . A composition comprising a fragment of TR2, wherein the composition interacts with ER, such that ER transcriptional activity is decreased relative to transcriptional activity in the absence of the composition, wherein the fragment of TR2 has at least 80%, 85%, 90%, or 95% identity to amino acids 88-196 of SEQ ID NO:10.  
     
     
         3 . The composition of  claim 2 , wherein any variation between the TR2 and the sequence set forth in SEQ ID NO: 10 is a considered a conserved variation.  
     
     
         4 . The composition of  claim 2 , wherein the composition reduces the transcription activity of ER.  
     
     
         5 . The composition of claims  2 , wherein the composition reduces the transcription activity of ER by 10%, 25%, 50%, or 90%.  
     
     
         6 . The composition of  claim 2 , wherein the composition reduces the G1/S transition of the cell cycle.  
     
     
         7 . The composition of claims  2 , wherein the composition reduces the G1/S transition of the cell cycle by 10%, 25%, 50%, or 90%.  
     
     
         8 . A method of inhibiting transcription activity of ER comprising administering the composition of  claim 1 .  
     
     
         9 . The method of  claim 8 , wherein the composition reduces the transcription activity of ER.  
     
     
         10 . The method of  claim 8 , wherein the composition reduces the transcription activity of ER by 10%, 25%,50%, or 90%.  
     
     
         11 . The method of  claim 8 , wherein the composition reduces the GI/S transition of the cell cycle.  
     
     
         12 . The method of  claim 8 , wherein the composition reduces the G1/S transition of the cell cycle by 10%, 25%, 50%, or 90%.  
     
     
         13 . A method of inhibiting TR2 transcription activity comprising administering a composition that binds TR2, wherein the composition is ER, ER fragment, ER variant, a molecule that competitively competes with TR2 for ER binding, or combination thereof.  
     
     
         14 . A method of identifying an inhibitor of an interaction between ER and TR2, comprising incubating a library of molecules with an ER, ER fragment, ER variant or combination, forming a mixture, and identifying the molecules that disrupt the interaction between the ER, ER fragment, ER variant, or combination and TR2, wherein the interaction disrupted comprises an interaction between the ER, ER fragment, ER variant, or combination and TR2 binding site.  
     
     
         15 . The method of  claim 14 , wherein the step of isolating comprises incubating the mixture with a molecule comprising TR2, TR2 fragment, TR2 variant, or combination.  
     
     
         16 . A method of identifying an inhibitor of an interaction between ER and TR2 comprising incubating a library of molecules with TR2, TR2 fragment, TR2 variant or combination forming a mixture, and identifying the molecules that disrupt the interaction between ER and the TR2, TR2 fragment, TR2 variant or combination, wherein the interaction disrupted comprises an interaction between the ER and the TR, TR2 fragment, TR2 variant or combination binding site.  
     
     
         17 . The method of  claim 16 , wherein the step of isolating comprises incubating the mixture with molecule comprising ER, ER fragment, ER variant, or combination.  
     
     
         18 . A composition comprising a fragment of ER, wherein the composition interacts with TR2, such that TR2 transcriptional activity is decreased relative to transcriptional activity in the absence of the composition, wherein the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to the sequence set forth in SEQ ID NO:32, the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 312-340, set forth in SEQ ID NO:32, the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 123-340, set forth in SEQ ID NO:32, or the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 312-595, set forth in SEQ ID NO:32.  
     
     
         19 . The composition of  claim 18 , wherein any variation between the ER and the sequence set forth in SEQ ID NO: 32 is a conserved variation.  
     
     
         20 . The composition of  claim 18 , wherein the fragment comprises the sequence set forth in SEQ ID NO:32, or is a conserved variant thereof.  
     
     
         21 . The composition of claims  18 , wherein the composition reduces the transcription activity of TR4.  
     
     
         22 . The composition of claims  18 , wherein the composition reduces the transcription activity of TR4 by 10%, 25%, 50%, or 90%.  
     
     
         23 . A method of inhibiting transcription activity of TR2 comprising administering the composition of claims  18 .  
     
     
         24 . The composition of claims  23 , wherein the composition reduces the transcription activity of TR4.  
     
     
         25 . The composition of claims  23 , wherein the composition reduces the transcription activity of TR4 by 10%, 25%, 50%, or 90%.  
     
     
         26 . A method of identifying inhibitors of ER transcription activity comprising mixing a compound with ER, ER fragment, ER variant, or combination and identifying compounds which compete with TR2 binding with ER, ER fragment, ER variant, or combination.  
     
     
         27 . A method of identifying inhibitors of ER transcription activity comprising mixing a set of compounds with TR2, TR2 fragment, TR variant, or combination and identifying compounds which compete with ER binding with TR2, TR2 fragment, TR variant, or combination.  
     
     
         28 . The method of claims  15 , wherein the TR comprises amino acids 88-196 of SEQ ID NO:10.  
     
     
         29 . The method of claims  28 , wherein the TR2 has a sequence of at least 80%, 85%, 90%, or 95% identity to amino acids 88-196 of SEQ ID NO:10.  
     
     
         30 . The method of claims  29 , wherein any variation between the TR2 and the sequence set forth in SEQ ID NO: 10 is considered a conserved variation.  
     
     
         31 . The method of claims  14 , wherein the identified compound binds ER with a k d  less than or equal to 10 −5  M, 10 −6  M, 10 −7  M, 10 −8  M, 10 −9  M, or 10 −10  M, 10 −11  M, or 10 −12  M.  
     
     
         32 . The method of claims  14 , wherein the ER comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to the sequence set forth in SEQ ID NO:32.  
     
     
         33 . The method of claims  14 , wherein the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 312-340, set forth in SEQ ID NO:32, the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 123-340, set forth in SEQ ID NO:32, or the fragment comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to amino acids 312-595, set forth in SEQ ID NO:32.  
     
     
         34 . The method of  claim 33 , wherein any variation between the ER and the sequence set forth in SEQ ID NO: 32 is a conserved variation.  
     
     
         35 . The method of  claim 33 , wherein the ER, ER fragment, ER variant or combination comprises the sequence set forth in SEQ ID NO:32, the sequence set forth by amino acids 312-340 of the sequence set forth in SEQ ID NO:32, the sequence set forth by amino acids 123-340 of the sequence set forth in SEQ ID NO:32, the sequence set forth by amino acids 312-595 of the sequence set forth in SEQ ID NO:32, or is a conserved variant thereof, or is a fragment thereof.  
     
     
         36 . A method of inhibiting TR4 transcription activity comprising administering a composition that binds TR4, wherein the composition is AR, AR fragment, AR variant, a molecule that competitively competes with TR4 for AR binding, or a combination thereof.  
     
     
         37 . A method of inhibiting AR transcription activity comprising administering a composition that binds AR, wherein the composition is TR4, TR4 fragment, TR4 variant, a molecule that competitively competes with AR for TR4 binding, or combination thereof.  
     
     
         38 . A method of identifying an inhibitor of an interaction between AR and TR4, comprising incubating a library of molecules with AR, AR fragment, or AR variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between AR, AR fragment, or AR variant, or combination and TR4, wherein the interaction disrupted comprises an interaction between the AR, AR fragment, or AR variant, or combination and TR4 binding site.  
     
     
         39 . The method of  claim 38 , wherein the step of isolating comprises incubating the mixture with molecule comprising TR4, TR4 fragment, TR4 variant or combination.  
     
     
         40 . A method of identifying an inhibitor of an interaction between AR and TR4 comprising incubating a library of molecules with TR4, TR4 fragment, or TR4 variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between AR and the TR4, TR4 fragment, or TR4 variant, or combination, wherein the interaction disrupted comprises an interaction between the AR and the TR4, TR4 fragment, or TR4 variant, or combination binding site.  
     
     
         41 . The method of  claim 42 , wherein the step of isolating comprises incubating the mixture with molecule comprising AR, AR fragment, AR variant or combination.  
     
     
         42 . The method of  claim 41 , wherein the AR, AR fragment, AR variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:31.  
     
     
         43 . The method of 42, wherein any variation between the AR and the sequence set forth in SEQ ID NO: 31 is a conserved variation.  
     
     
         44 . The method of  claim 36 , wherein the transcription activity of TR4 is reduced by 10%, 25%, 50%, or 90%.  
     
     
         45 . The method of  claim 40 , wherein the TR4, TR4 fragment, TR4 variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:16.  
     
     
         46 . The method of 45, wherein any variation between the TR4 and the sequence set forth in SEQ ID NO: 16 is a conserved variation.  
     
     
         47 . The method of  claim 37 , wherein the transcription activity of AR is reduced by 10%, 25%, 50%, or 90%.  
     
     
         48 . A method of inhibiting TR4 transcription activity comprising administering a composition that binds TR4, wherein the composition is ER, ER fragment, ER variant, or a molecule that competitively competes with TR4 for ER binding or combination thereof.  
     
     
         49 . A method of inhibiting ER transcription activity comprising administering a composition that binds ER, wherein the composition is TR4, TR4 fragment, TR4 variant, a molecule that competitively competes with ER for TR4 binding, or combination thereof.  
     
     
         50 . A method of identifying an inhibitor of an interaction between ER and TR4, comprising incubating a library of molecules with ER, ER fragment, ER variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between the ER, ER fragment, ER variant, or combination and TR4, wherein the interaction disrupted comprises an interaction between the ER, ER fragment, ER variant, or combination and TR4 binding site.  
     
     
         51 . The method of  claim 50 , wherein the step of isolating comprises incubating the mixture with molecule comprising TR4, TR4 fragment, TR4 variant or combination.  
     
     
         52 . A method of identifying an inhibitor of an interaction between ER and TR4 comprising incubating a library of molecules with TR4, TR4 fragment, TR4 variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between ER and TR4, TR4 fragment, TR4 variant, or combination, wherein the interaction disrupted comprises an interaction between the ER and TR4, TR4 fragment, TR4 variant, or combination binding site.  
     
     
         53 . The method of  claim 51 , wherein the step of isolating comprises incubating the mixture with a molecule comprising an ER, ER fragment, ER variant or combination.  
     
     
         54 . The method of  claim 53 , wherein the ER, ER fragment, ER variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:32.  
     
     
         55 . The method of 54, wherein any variation between the ER and the sequence set forth in SEQ ID NO: 32 is a conserved variation.  
     
     
         56 . The method of  claim 48 , wherein the transcription activity of TR4 is reduced by 10%, 25%, 50%, or 90%.  
     
     
         57 . The method of  claim 52 , wherein the TR4, TR4 fragment, TR4 variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:16.  
     
     
         58 . The method of 57, wherein any variation between the TR4 and the sequence set forth in SEQ ID NO: 16 is a conserved variation.  
     
     
         59 . The method of  claim 49 , wherein the transcription activity of ER is reduced by 10%, 25%, 50%, or 90%.  
     
     
         60 . A method of inhibiting AR transcription activity comprising administering a composition that binds AR, wherein the composition is TR2, TR2 fragment, TR2 variant, a molecule that competitively competes with AR for TR2 binding or combination thereof.  
     
     
         61 . A method of identifying an inhibitor of AR transcription activity comprising mixing a compound with AR, AR fragment, AR variant, or combination and identifying compounds which compete with the TR2 interaction with AR, AR fragment, AR variant, or combination.  
     
     
         62 . A method of identifying inhibitors of AR transcription activity comprising mixing a set of compounds with TR2, TR2 fragment, TR2 variant or combination thereof and identifying compounds which compete with TR2, TR2 fragment, TR2 variant or combination interaction with AR.  
     
     
         63 . A method of identifying an inhibitor of an interaction between AR and TR2, comprising incubating a library of molecules with AR, AR fragment, AR variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between AR, AR fragment, AR variant, or combination and TR2, wherein the interaction disrupted comprises an interaction between the AR, AR fragment, AR variant, or combination and TR2 binding site.  
     
     
         64 . The method of  claim 62 , wherein the step of isolating comprises incubating the mixture with molecule comprising TR2, TR2 fragment, TR2 variant or combination.  
     
     
         65 . A method of identifying an inhibitor of an interaction between AR and TR2 comprising incubating a library of molecules with TR2, TR2 fragment, TR2 variant, or combination forming a mixture, and identifying the molecules that disrupt the interaction between AR and the TR2, TR2 fragment, TR2 variant, or combination, wherein the interaction disrupted comprises an interaction between the AR and the TR2, TR2 fragment, TR2 variant, or combination binding site.  
     
     
         66 . The method of  claim 65 , wherein the step of isolating comprises incubating the mixture with molecule comprising AR, AR fragment, AR variant or combination.  
     
     
         67 . The method of  claim 63 , wherein the AR, AR fragment, AR variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:31.  
     
     
         68 . The method of  67 , wherein any variation between the AR and the sequence set forth in SEQ ID NO:31 is a conserved variation.  
     
     
         69 . The method of  claim 68 , wherein the transcription activity of TR2 is reduced by 10%, 25%, 50%, or 90%.  
     
     
         70 . The method of  claim 65 , wherein the TR2, TR2 fragment, TR2 variant or combination comprises a polypeptide having at least 80%, 85%, 90%, or 95% identity to SEQ ID NO:16.  
     
     
         71 . The method of 70, wherein any variation between the TR2 and the sequence set forth in SEQ ID NO:16 is a conserved variation.  
     
     
         72 . The method of  claim 60 , wherein the transcription activity of AR is reduced by 10%, 25%, 50%, or 90%.  
     
     
         73 . An isolated composition comprising TR2 and ER or variants or fragments of either, TR2 and AR or variants or fragments of either, TR4 and ER or variants or fragments of either, or TR4 and AR or variants or fragments of either.